Figure 4
From: Jedi-1 deficiency increases sensory neuron excitability through a non-cell autonomous mechanism
DRG neurons isolated from Jedi-1 KO mice are sensitized to capsaicin. (A) Representative trace of 4 cells used to obtain live cell calcium imaging data. Duration of treatment with allylisothiocyanate (AITC), capsaicin (CAP), and KCl is shown in grey boxes. Blue cell is a healthy neuron that responded to KCl but not AITC or CAP. Red cell responded to AITC and KCl. Purple cell responded to CAP and KCl. CAP responding cells often did not return to baseline. Green cell responded to AITC, CAP, and KCl. (B) Venn diagrams showing the proportions of cells that responded to AITC and CAP. t tests were used to compare overall responses to each drug. There was no statistical difference in AITC responses between genotypes. (C) Fraction of cells that responded to capsaicin over the course of 3 independent experiments with a minimum of 100 cells analyzed in each experiment. Two-tailed t test with Welch’s correction p = 0.03. (D) Intracellular calcium concentration measured by Fura-2AM. 34. Baseline calcium concentration was averaged over the course of 1 minute before any drug treatment. Maximum calcium concentration is the peak calcium concentration during treatment with 500 nM capsaicin for 30 seconds. Each data point represents an independent experiment with a minimum of 100 cells analyzed in each. Students t test was used to compare calcium concentrations between genotypes at baseline and after CAP treatment. (E) Left: Quantification of the size of of TrpV1+ neurons in vitro. n = 4 animals per genotype analyzed. Scale bars represent SEM. 2 way ANOVA used to compare genotypes across all size categories. No statistical differences between genotypes. Right: Representative image of TrpV1 staining in primary DRG cultures showing neurons in Tuj1 (green) and TrpV1 (red). All images were analyzed in ImageJ version 2.0.0-rc-69/1.52p.
